Composition based on xyloglucan and proteins for the treatment of intestinal disorders

ABSTRACT

Disclosed are compositions comprising synergic combinations of xyloglucans and plant or animal proteins, which are useful in the treatment of intestinal disorders.

This Non-Provisional application is a continuation of U.S. Ser. No.15/597,664, filed on May 17, 2017, which is a continuation of U.S. Ser.No. 15/303,753, filed on Oct. 13, 2016, now abandoned, which is a U.S.National Stage of PCT/EP2015/058162 filed on 15 Apr. 2015, which claimspriority to and the benefit of Italian Application No. MI2014A000705filed on 15 Apr. 2014, the contents of which are incorporated herein byreference in their entireties.

The invention relates to synergic combinations of xyloglucans and plantor animal proteins and compositions for the treatment of intestinaldisorders, especially diarrhoeal forms of various origins.

PRIOR ART

Diarrhoea is a symptom of many gastrointestinal disorders and is oftenincapacitating and dangerous, especially in children and the elderly.Acute diarrhoea is mainly caused by intestinal infections, but can alsobe due to the use of medicaments or radiotherapy and to otherpathological conditions (diverticulitis, heavy-metal poisoning,intestinal ischaemia, allergies and intolerances).

Acute diarrhoea with an infectious cause is a serious problem indeveloping countries; it is believed to cause the death of at least 4million children under 5 years old every year.

Chronic diarrhoea is generally due to irritable bowel syndrome, coeliacdisease or inflammatory bowel diseases (Crohn's disease, ulcerativerectocolitis).

In view of their different aetiologies, various treatment options areavailable, based on the administration of antibiotics/antibacterials,spasmolytics/anticholinergics, probiotics, or opioid receptor agonists.However, some of said treatments must be administered with greatcaution, because they do not act on the causal pathological process.

To prevent said adverse effects, complexes of tannins complexed withanimal proteins and gelatins, in particular with gelatin of bovineorigin, albumin, casein or ovalbumin, have been proposed for some time.

For example, the use of said complexes in the treatment of the variousforms of diarrhoea is disclosed in EP 1764105, EP 2526939, EP 2361623and US 20090062191. Gelatin tannate has been available on the market forsome time as a medical device for the treatment of acute diarrhoea.

Xyloglucans are molecules consisting of a linear backbone ofβ-1,4-glucans with short side branches. The latter bond due to thexylose bonded to oxygen in the 6 position of the sugar. Said side chainscan also contain other sugars such as arabinose and fucose.

Xyloglucans belong to the hemicellulose family, which combines withcellulose in the cell wall of the higher plants. A particularly richsource of xyloglucan is the seeds of tamarind (Tamarindus indica), atropical tree originating from East Africa.

Xyloglucan-rich tamarind seed extracts are known and have been used inthe medical field mainly as viscosity-controlling agents in ophthalmiccompositions (U.S. Pat. No. 6,056,950), as mucoadhesive agents (WO2006131262), as artificial tears (WO 2009/044423), as anti-infectiveagents (WO 2011147767) and as anti-inflammatory agents (WO 2011147768).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows that xyloglucan alone did not reduce the fluid secretion.

DESCRIPTION OF THE INVENTION

It has now surprisingly been found that combinations of xyloglucans withplant or animal proteins compatible with oral administration to humansare particularly effective in the treatment and prevention of diarrhoeaand other infectious and/or inflammatory intestinal disorders.Xyloglucans exert a film-forming effect in the intestinal mucosa whichreduces the permeability of the tight junctions of the intestinalmucosa, and therefore prevents the entry of the pathogens responsiblefor acute intestinal infections. The film-forming effect is not affectedby variations in pH.

The invention therefore relates to pharmaceutical compositionscomprising, as active ingredients, xyloglucans or extracts containingthem, combined with at least one plant or animal protein selected fromgelatine, albumin, ovalbumin, casein, pea protein and soya protein andsuitable excipients, and optionally with other active ingredients usefulfor the prevention and treatment of gastrointestinal and urogenitaldisorders.

Xyloglucans extracted from Tamarindus indica are available on themarket, for example from Indena (Italy) (Xilogel®) and DSP Gokyo Food &Chemical (Japan) (Glyloid®). The average molecular weight is between400,000 and 650,000 daltons.

Preferred proteins include gelatin and pea protein. Gelatin isparticularly preferred.

The weight ratio of xyloglucan to protein ranges between 1:0.5 and 1:30.The combination of xyloglucan and protein forming the subject of theinvention constitutes the active ingredient of oral pharmaceuticalformulations.

Examples of suitable forms of administration include capsules, tablets,solutions, suspensions, granules, gels and the like.

Other active ingredients with which xyloglucans and protein can becombined include antibiotics, antimotility agents, steroidal andnon-steroidal anti-inflammatories, compounds for the treatment ofgastrointestinal bloating (simethicone and the like), mesalazine,sucralfate, natural and synthetic polysaccharides such as pectins,chitosan (animal or vegetable), hyaluronic acid, guar gum, xanthan gum,cellulose and hemicellulose and derivatives such ashydroxypropylcellulose, carrageenans, carbomers, andcrosslinking/polymerising compounds such as ferulic acid; polyphenols,such as gall polyphenols, polyphenols from grape pips, probiotics suchas Lactobacilli, Bifidobacteria, yeasts and the like.

In the compositions according to the invention, xyloglucans can bepresent in a wide concentration range which depends on the type ofcomposition and the therapeutic indication for which they are intended.

The xyloglucan is administered orally at doses ranging between 0.5mg/dose and 200 mg/dose, preferably between 10 mg/dose and 100 mg/dose.

The protein, in particular gelatin, is administered orally at dosesranging between 10 mg/dose and 3000 mg/dose, preferably between 50mg/dose and 500 mg/dose.

The compositions according to the invention are useful for the treatmentand prevention of gastrointestinal disorders and other disorders thatoriginate in the gastrointestinal system and are transferred to othersystems, such as the urogenital system. It is known that theGram-negative bacteria present in the intestine, in particularEscherichia coli, can proliferate in said organ and migrate to theurinary tract, where they cause 90% of urogenital infections such ascystitis, cystopyelitis and the like.

In particular, the compositions according to the invention are useful toprevent the proliferation of pathogens in the gastrointestinal systemand transfer them to other systems of the human body through the tightintestinal junctions, to protect the intestinal mucosa against chemicalor physical agents which can reduce the functionality and naturalregeneration of the intestinal epithelium, and to reduce theparacellular flow of pathogens through the intestinal walls.

The compositions according to the invention have also proved useful forthe prevention and treatment of damage to the intestinal mucosa and theconsequent inflammatory symptoms, such as diverticulosis and the earlystages of diverticulitis; for the treatment of symptoms resulting fromfood allergies (e.g. intolerance of lactose, gluten, etc.); for theprevention and treatment of digestive disorders (gas production,bloating, stomach rumble and flatulence); and for the prevention andtreatment of damage to the intestinal mucosa deriving from localinflammatory phenomena of transient or chronic origin, in particular forthe treatment of Crohn's disease, ulcerative colitis and irritable bowelsyndrome (IBS).

The compositions according to the invention can be advantageously usedto treat diarrhoea in combination with oral rehydration electrolytes,such as mucomimetics, and to inhibit the adherence of bacteria to themucosa and subsequent proliferation involving dysbiosis, optionallycombined with probiotics or tyndallised bacteria. The compositionsaccording to the invention are useful for the prevention and treatmentof travellers' diarrhoea.

The compositions according to the invention effectively protect themucosa and reduce the adherence to it of some pathogens, such asgas-producing bacteria.

The examples below illustrate the invention in greater detail.

Example 1

Composition for the Prevention and Treatment of Diarrhoea; Single-DoseSachet

Xyloglucan 0.100 g Gelatin 0.050 g Inulin 1.650 g Maltodextrin 1.195 gStevioside (Stevia) 0.015 g Tuttifrutti flavouring (Firmenich) 0.015 gE160 (a) colouring (betacarotene) 0.025 g

Example 2

Composition for the Prevention and Treatment of Diarrhoea; Hard Capsule

Xyloglucan 0.1 g Gelatin 3.0 g Matricaria E.S. 0.026 g Pectin 0.050 gDimethicone 0.020 g Kaolin 0.020 g Magnesium stearate 0.080 g

Example 3

Composition for the Prevention and Treatment of Diarrhoea; Tablet

Xyloglucan 0.1 g Pea protein 0.5 g Lactose 0.063 g Anhydrous colloidalsilicon dioxide 0.002 g Microcrystalline cellulose 0.030 g Magnesiumstearate 0.003 g

Example 4—Bioassays: Protection Against Intestinal Fluid SecretionInduced by Cholera Toxin in Rats

Four groups of Wistar rats (200-220 g) were treated orally with 12.5mg/kg of xyloglucan, 125 mg/kg of gelatin and the combination of saidtwo ingredients of the combination, at the same dose. Six hours afteradministration, the groups of animals were treated with cholera toxin atthe dose of 6 μg/ml.

Two hours after the toxin treatment the water content of the intestinalloop was measured.

The results obtained, shown in the FIG. 1 and in the following Table,demonstrate that xyloglucan alone did not reduce the fluid secretion.Equally, gelatin alone did not exhibit a significant effect, whereas theeffects of the combination proved statistically significant.

12.5 mg 12.5 mg 125 mg 12.5 mg xyloglucan/kg/PO⁵ + xyloglucan/kg/PO⁵ +Saline + gelatine/kg/PO³ - xyloglucan/kg/PO⁷ - Gelatin (125 mg/kg)⁶ -Gelatin (250 mg/kg)⁶ - Basal ¹ CT² 6 hours 6 hours 6 hours 12 hoursGrams/loop 0.41 ± 0.11 1.04 ± 0.32 1.01 ± 0.39 1.26 ± 0.18 0.77 ± 0.150.75 ± 0.16 p NS⁴ NS⁴ NS Significant Significant (p < 0.01) (p < 0.05)

Example 5—Clinical Trial

A multicentre controlled parallel-group clinical trial was conducted byadministering to patients suffering from acute diarrhoea the combinationaccording to the invention (xyloglucan 400 mg/day and gelatin 200mg/day), the probiotic S. boulardii (at the dose of 7×10⁹ cells/dose),and diosmectite (Smecta®, 3×3 g sachets/day). The speed of onset ofclinical efficacy was evaluated in the three groups (reduction induration of acute diarrhoea and the correlated symptoms). The symptomsexamined were nausea, vomiting, flatulence, abdominal pain and stoolemissions. The symptoms declined in all three groups. The combinationaccording to the invention led to more rapid action, inhibiting thediarrhoea within 24 hours of the start of the treatment. Abdominal painwas monitored throughout the treatment. The patients did not presentvomiting after 48 and 72 hours. The combination according to theinvention gave rise to a more rapid reduction in stool emissions ratedas grades 6 and 7 on the Bristol scale, with a 60% reduction as against34% and 39% respectively for diosmectite and S. boulardii. After 48hours this type of emission had almost entirely disappeared in all threegroups. The combination according to the invention therefore proved tobe the fastest-acting in preventing stool emissions.

The invention claimed is:
 1. Method of treating diarrhoea, Crohn'sdisease, ulcerative colitis and irritable bowel syndrome (IBS),diverticulosis, coeliac disease, lactose intolerance, cystitis andcystopyelitis in humans in need thereof, said method comprisingadministering to said humans an effective amount of a combination ofxyloglucans or extracts containing them and pea protein, wherein theweight ratio between xyloglucan and pea protein is 1:0.5 and 1:30. 2.The method according to claim 1, wherein the weight ratio betweenxyloglucan and pea protein is 1:5.
 3. The method according to claim 1further comprising excipients.
 4. The method according to claim 3further comprising additional active ingredients selected from the groupconsisting of antibiotics, antimotility agents, steroidal ornon-steroidal anti-inflammatories, compounds for the treatment ofgastrointestinal bloating, natural or synthetic polysaccharides, andelectrolytes.
 5. The method according to claim 4, wherein saidnon-steroidal anti-inflammatories agents and said naturalpolysaccharides are selected from the group consisting of mesalazine,sucralfate, hyaluronic acid, guar gum, xanthan gum, cellulose andhemicellulose, hydroxypropyl cellulose, carrageenans, carbomers, ferulicacid; polyphenols and probiotics.